Substituted quinolines as noncovalent proteasome inhibitors

Article abstract of DOI:10.1016/j.bmc.2016.04.005

Screening of a library of diverse heterocyclic scaffolds identified substituted quinolines as inhibitors of the human proteasome. The heterocyclic library was prepared via a novel titanium-catalyzed multicomponent coupling reaction, which rendered a diver

Full text of DOI:10.1016/j.bmc.2016.04.005

Accepted Manuscript
Substituted Quinolines as Noncovalent Proteasome Inhibitors
Tanner J. McDaniel, Theresa A. Lansdell, Amila A. Dissanayake, Lauren M.
Azevedo, Jacob Claes, Aaron L. Odom, Jetze J. Tepe
PII:
S0968-0896(16)30239-5
DOI:
http://dx.doi.org/10.1016/j.bmc.2016.04.005
BMC 12919
Reference:
To appear in:
Bioorganic & Medicinal Chemistry
Received Date:
Revised Date:
Accepted Date:
18 December 2015
23 March 2016
1 April 2016
Please cite this article as: McDaniel, T.J., Lansdell, T.A., Dissanayake, A.A., Azevedo, L.M., Claes, J., Odom, A.L.,
Tepe, J.J., Substituted Quinolines as Noncovalent Proteasome Inhibitors, Bioorganic & Medicinal Chemistry (2016),
doi: http://dx.doi.org/10.1016/j.bmc.2016.04.005
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers
we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and
review of the resulting proof before it is published in its final form. Please note that during the production process
errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Graphical Abstract
To create your abstract, type over the instructions in the template box below.
Fonts or abstract dimensions should not be changed or altered.
Substituted Quinolines as Noncovalent
Leave this area blank for abstract info.
Proteasome Inhibitors
Tanner J. McDaniel, Theresa A. Lansdell, Amila A. Dissanayake, Lauren M. Azevedo, Jacob Claes, Aaron L. Odom* and Jetze J. Tepe*

Bioorganic & Medicinal Chemistry
Substituted Quinolines as Noncovalent Proteasome Inhibitors
Tanner J. McDaniel, Theresa A. Lansdell, Amila A. Dissanayake, Lauren M. Azevedo, Jacob Claes, Aaron L. Odom* and Jetze J. Tepe*
Department of Chemistry, Michigan State University, East Lansing, MI 48824
ARTICLE INFO
ABSTRACT
Article history:
Received
Revised
Accepted
Available online
Screening of a library of diverse heterocyclic scaffolds identified substituted
quinolines as inhibitors of the human proteasome. The heterocyclic library was
prepared via a novel titanium-catalyzed multicomponent coupling reaction, which
rendered a diverse set of isoxazoles, pyrimidines, pyrroles, pyrazoles and quinolines.
SAR of the parent lead compound indicated that hydrophobic residues on the benzo-
moiety significantly improved potency. Lead compound 25 inhibits the chymotrypic-
like proteolytic activity of the proteasome (IC50 5.4M), representing a new class of
nonpeptidic, noncovalent proteasome inhibitors.
Keywords:
Proteasome
Inhibitors
Noncovalent
Quinolines
@
2015 Elsevier Ltd. All rights reserved.
1
.
Introduction
These suicide inhibitors effectively block global protein
proteolysis, which induces apoptosis, but also triggers a
transcriptional feedback loop that results in the synthesis of new
Proteins undergo constant proteolytic degradation to regulate
intracellular processes and maintain biological homeostasis.
During this process, redundant and misfolded proteins are tagged
with ubiquitin, which marks them for proteolytic degradation by
the 26S proteasome. The 26S proteasome is comprised of a barrel
shaped 20S core particle (CP) that is capped by two 19S
recognition particles (RPs). The 20S CP is a threonine protease
that contains three distinct catalytic subunits (5, 2 and 1) that
exhibit chymotrypsin-like (CT-L), trypsin-like (T-L) and
1
,2
21
proteasome subunits. In addition, peptidase cleavage induces
10,22
the rapid systemic clearance of these inhibitors.
Although the
initial burst of inhibition is highly effective in the induction of
apoptosis, the unfavorable pharmacodynamic properties of these
peptide-based suicide inhibitors have restricted their use to blood
cancers.
3
Noncovalent and nonpeptidic proteasome inhibition may limit
4
caspase-like (Casp-L) activity.
recognizes the ubiquitin tagged proteins and is responsible for the
unfolding and translocation of these substrates into the 20S
The 19S regulatory particle
some of these intrinsic pharmacokinetic drawbacks and may
23,24
translate into a broader clinical profile.
Relative to covalent
binders, reports of noncovalent and nonpeptidic proteasome
inhibitors are still scarce, but are gaining recognition as viable
5
proteolytic core chamber.
11
Modulation of proteasome function has emerged as an
alternatives to peptide-based suicide inhibitors.
6,7
important approach to treat various diseases, and several
proteasome inhibitors are clinically approved.
8-10
Common
themes among proteasome inhibitors are that they are protein
mimics comprised of a peptide backbone containing an
1
1-13
electrophilic warhead.
The warhead forms a covalent bond
1
with the N terminal Thr in the catalytic site(s) of the 20S core
particle and abrogates its enzymatic activity. Thus, all these
peptide based suicide inhibitors are competitive inhibitors that
bind in the catalytic site(s) of the proteasome. The peptide-
based suicide inhibitors bortezomib (Fig. 1, BTZ or Velcade) and
carfilzomib (CFZ, Kyprolis) are FDA approved for the treatment
11
14-17
of multiple myeloma (MM)
and mantle cell lymphoma
(
MCL) and have validated the proteasome as an important
18-20
clinical target.
Figure 1. Structures of covalent, peptidic proteasome inhibitors bortezomib,
and noncovalent, nonpeptidic proteasome inhibitors PI-083, Chloroquine,
5
AHQ and substituted quinolines.
*Corresponding authors: Tel: 517-355-9715; E-mail address:
Odom@chemistry.msu.edu & Tepe@chemistry.msu.edu

Examples of nonpeptidic noncovalent proteasome modulators
acid, which rendered highly substituted frameworks in a one-pot
procedure (Fig. 2).
25
26
24
43
include the phakellins,
oxadiazoles,
hydroxyureas,
27,28
29
imidazolines
sulfone or piperazine agents, and tamoxifen
The search for proteasome inhibition quickly narrowed to
compounds containing the quinoline backbone. Due to the
mechanism of the formation of the iminoamination products, the
quinolines are all unsubstituted at the 4-position but can be
substituted by a range of groups in other sites. The two catalysts
employed for the syntheses are shown at the bottom of figure 2.
3
0
derivatives.
Substituted quinolines in particular represent a very
interesting new class of proteasome inhibitors. Lawrence and co-
workers discovered and optimized
a
novel class of
hydrophthoquinone derivates as nonpeptidic, noncovalent
3
1,32
The ancillary ligands for titanium H dpma and H dpm are both
proteasome inhibitors.
Fig. 1), demonstrated selectivity for cancer cells over non-
transformed cells, which potentially broadens the range of
These agents, exemplified by PI-083
2
2
44-46
prepared in a single step from pyrrole.
The catalysts for this
(
study were isolated as pure compounds before use; however, it is
possible to generate the catalysts in situ from the protio-ligand
33
anticancer activity. Recently, crystallographic screening of
compounds revealed a sulfonamide substituted quinoline as a
noncovalent inhibitor of only the 1/2 subunits and not the 5
chymotryptic activity, thus identifying a new binding motif.
Several quinolines have also been found to allosterically
modulate proteasome activity. NMR studies found that the
structurally similar anti-malaria drug chloroquine was found to
allosterically modulate proteasome activity by binding to the /
interface of the Thermoplasma proteasome. Similarly, 5-amino-
-hydroxyquinoline (5-AHQ, Fig. 1) was reported to be a non-
40
and commercially available Ti(NMe ) as well. The applications
2
4
of these two catalysts in iminoamination have been discussed
4
0
3
4
elsewhere in detail. In short, more reactive Ti(dpm)(NMe ) is
2 2
often used for more difficult internal alkyne substrates, while
milder Ti(dpma)(NMe is often used with sensitive terminal
3
5
)
2 2
alkynes to avoid potential side reactions. In addition, the two
catalysts can direct regioselectivity for the substrates, which
broadens the structure diversity.
3
6
8
3
7
competitive inhibitor of the human proteasome. Importantly, 5-
AHQ was found effective against bortezomib resistant cell lines,
thus exemplifying another advantage of using mechanistically
2
7,38,39
distinct classes of proteasome inhibitors.
It is therefore
tempting to speculate that at least some of these noncovalent,
nonpeptidic quinolines may occupy a common allosteric binding
35
site.
In our search for noncovalent nonpeptidic proteasome
inhibitors, we screened several different classes of heterocyclic
scaffolds for their ability to inhibit the chymotryptic activity of
the human proteasome. The small diverse library was comprised
of various aminocyanopyridines, isoxazoles, pyrazoles and
quinolines that were prepared via
a
titanium-mediated
multicomponent coupling reaction (Fig. 2). Of the compounds
tested, only the quinolines exhibited low micromolar efficacies
for 20S proteasome inhibition. Following the identification of the
quinolines as initial hits in the in vitro screen, we characterized
its mechanism as a mixed-type inhibition of proteasome
modulation, which translated well in cell culture as indicated by
the reduction of NF-B mediated gene expression in a NF-B-luc
reporter assay in HeLa cells. In order to optimize the scaffolds
1 5
activity, we modified five points of diversity (Fig. 1, R -R )
along the scaffold backbone using the titanium catalyzed
multicomponent coupling reaction.
Figure 2. Titanium catalyzed multicomponent coupling reaction to form
pyridines, isoxazoles, pyrazoles, and quinolines, as examples.
Herein, we report the synthesis of a novel class of quinoline-
based scaffolds and their biological activity towards to 20S
proteasome. Importantly, these scaffolds are structurally distinct
from the hydro-phthoquinone and 5-AHQ, in that they do not
contain, or can readily generate, an electrophilic benzoquinone
moiety. In addition, the binding motif is likely different from the
The synthesis of some of the quinolines follows a similar
43
one-pot synthesis as previously reported. Synthetic details for
all new compounds can be found in the Supporting Information.
34
2.2. Biological evaluation
sulfonamide-based quinolines, as the compounds described
herein are potent inhibitors of chymotryptic-like activity.
The initial diverse library of compounds was screened in
vitro using purified human 20S proteasome and the fluorogenic
peptide substrate, Suc-LLVY-AMC, as the substrate for CT-L
2
2
.
Results and discussion
4
7
activity.
The rates of hydrolysis were monitored by
.1. Chemistry
fluorescence increase at 37° C over 30 minutes and the linear
portion of the curves were used to calculate the IC50 values. Of
the compounds tested, only some of the quinolines exhibited low
micromolar efficacies for 20S proteasome inhibition (Figure S1).
Of the quinolines tested, quinoline 7 exhibited modest inhibition
of the 20S chymotryptic activity with an IC50 of 14.4 M and was
therefore selected for further evaluation and optimization.
A novel titanium-catalyzed 3-component coupling reaction
was applied to generate a small library of diverse heterocyclic
40-42
scaffolds.
The titanium chemistry effectively adds an iminyl
and amine group across the triple bond of an alkyne,
iminoamination, to form unsymmetrical derivatives of 1,3-
diimines. The 1,3-diimines, generated in situ, can then be
applied to many different heterocyclic syntheses.
quinolines were prepared from the 3-component coupling
products using a modified Combes synthesis catalyzed by acetic
40
The
Interestingly, it appeared that substitutions in the R
positions were required to see any inhibition of proteasome
activity (Table 1).
1 2 3
, R , R and
R
5

Quinoline 7 was subsequently evaluated for its inhibition of
the proteasome’s tryptic (2)-like and caspase (1)-like activity
in vitro using purified human 20S proteasome and the following
incrementally and measurements were taken at five different
concentrations of compound 7 or vehicle, the Vmax of the CT-L
M
activity decreases and the K increases, with the increasing
concentration of substrate (Fig. 3B). This is a pattern that is
4
8
consistent with mixed-type inhibition, and is consistent with an
allosteric-type modulation of proteasome activity, where binding
of the quinoline 7 occurs at a site different from the active site,
resulting in inhibition of enzyme activity.
Table 1. Structure and IC50 values of substituted quinones for inhibition of
chymotrypic activity of the 20S proteasome. All IC50 values are averages of
two independent experiments (each performed in triplicate).
In order to evaluate whether the inhibition of proteasome
activity translated in cell culture, we evaluated compound 7 for
inhibition of NF-B regulation. Inhibition of proteasome, affects
multiple critical signaling pathways and the anti-cancer activity
of proteasome inhibitors has been linked, in part, to their ability
to inhibit the pro-inflammatory, anti-apoptotic NF-B signaling
I C 5 0
(  M )
R 1
-
R 2
-
R 3
-
R 4
-
R 5
-
B T Z
F I G . 1 )
0 . 0 0 6 2
  0 . 0 0 0 3 
4
9-51
(
pathway.
The nuclear transcription factor, NF-B is

sequestered in the cytoplasm by the inhibitory protein B, termed
IB. Activation of the NF-B pathway by cytokines, such as
TNF-, results in the rapid ubiquitinylation and proteasomal
degradation of IB, which releases NF-B for nuclear
1
C H
3
H
H
H
H
   

2
3
H
H
C H
3
H
H
H
H
H
H
   
52
translocation and gene transcription. Proteasome inhibitors
prevent IB from proteolytic degrading and result in an
accumulation of cytosolic ubiquitinylated IB following TNF-

   
5
3
activation. In order to determine if quinoline 7 affects NF-B
mediated gene transcription, we evaluated quinoline 7 in HeLa
NF-B-luc cells.

4
5
6
H
C H
H
3
H
CH3
H
C H
H
3
    
     


CH
3



150
CH
3
CH
3
CH
3
15.3
±2.4)
100
(
*

50
7
C H
3
C H
3
H
C H
3
    

     
*
*
0
0.0
0.5
1.0
1.5
2.0
Log (M compound 7)
fluorogenic peptide as substrates: Boc-LRR-AMC (substrate for
T-L activity) and Z- LLE-AMC (substrate for casp-L activity).
The data shows that the quinoline 7 inhibits the casp-L at a
similar IC50 (IC50 17.7 M) as the chymotryptic activity, but not
the tryptic-L activities of the 20S catalytic core (IC50 >25 M,
Fig. 3).
47
Figure 4. NF-B-luc HeLa cells were unstimulated/stimulated with 25 ng/mL
TNF- in the absence or presence of quinoline 7 in 1% DMSO. Fold-
induction (%) of the Luciferase activity, normalized for all samples to TNF-
stimulation, is shown. Data shown is an average of 2 independent
experiments. * Statistically significant from the vehicle + TNF control (one-
way ANOVA n=4).
A
B
Cell cultures were pretreated with vehicle (1% DMSO), the
proteasome inhibitor MG-132 (positive control) or compound 7
(final concentrations were 50, 25, 12.5, 6.25, 3.13, 1.56 M) for
CT-L
casp-L
T-L
1
50
3
0 minutes at 37 ºC in 5% CO
concentration of 25 ng/mL, and the samples were further
incubated for 8 hours at 37 ºC in 5% CO and subsequently
2
. TNF- was added to a final
100
2
5
0
assayed for firefly luciferase production using the Steady-Glo
luciferase reporter assay. The in vitro inhibition of the
proteasome by quinoline 7 translated well in cell culture and
prevented NF-B mediated gene transcription with an IC50 value
of 12.1 M (Fig. 4) in a dose-response manner.
Following the cellular validation of the observed in vitro
proteasome activity, we moved to improve the potency and gain
insight into the structural requirements for activity. For this, we
expanded the scope of the 3-component coupling reaction to
include several addition functional groups. Using the same assay
conditions, several additional quinolines were evaluated for their
ability to inhibit the chymotrypic activity of the proteasome.
0
-
1.0
-0.5
0.0
0.5
1.0
1.5
Log (M quinoline 7)
Figure 3. A. Quinoline 7 inhibits the CT-L and Casp-L activity of the human
proteasome. Fluorogenic substrates Suc-LLVY-AMC, Z-ARR-AMC and Z-
LLE-AMC were used to measure CT-L, T-L and Casp-L activities of purified
human 20S proteasome particles. The maximum increase in fluorescence per
minute was used to calculate specific activities of each sample. B.
Lineweaver-Burk plot using SucLLVY-AMC consistent with a model of
“
mixed-type inhibition”. Undetectable proteasome activity was observed at
the lowest amount of substrate thus these values are omitted in the L-B plot.
The mechanism by which quinoline 7 inhibits the proteasome
was investigated using Michaelis-Menton analysis to determine
of K and Vmax and then further illustrated using a Lineweaver-
M
Burk double reciprocal plot of the kinetic data. Kinetic analysis
of CT-L activity of purified 20S particles indicate that when the
substrate (Suc-LLVY-AMC) concentration was increased
First, we examined some changes in the R
1
position (Table 2,
7
-9), which rendered minor improvements in potency.
Replacement of the cyclohexene with an isobutylene group (10),
rendered an inactive compound, indicating the significance of the
cyclohexene moiety. A significant gain (2-fold) in potency was

seen using the cyclohexyl moiety (11), which provided our first
single digit microMolar proteasome inhibitor. Activity decreased
using a phenyl group in this position (Table 1, compound 6). A
2
2
2
3
CH
3
H
H
N(CH
3
3
)
2
2
H
H
6.1
( ±0.2)
CH
2
CH
3
N(CH
)
5.6
± 0.4)
1
significant drop in activity was seen by replacing the R group of
(
compound 6 with a hydrogen (Table 1, 4) or aryl moiety (12)
indicating the need for small hydrophobic moieties around the
quinoline scaffold.
2
2
4
5
CH
3
3
H
H
H
H
9.1
±0.5)
(
CH
5.4
±0.1)
Next we evaluated the R
motif and compared these to our original lead (7), which
contained a methyl in the R and the cyclohexene ring in the R
position. Eliminating the R and R methyl groups, rendered
3 5
-R positions around the quinoline
(
2
2
6
7
CH
3
3
H
OCH
3
H
>25.0
1
2
3
5
CH
OCH
3
OCH
3
OCH
3
15.6
±0.7)
inactive compounds (13), indicating the need of hydrophobic
groups in these domains. Not even the lipophilic chloride (14) or
(
2
2
8
9
H
H
H
N(CH
3
)
2
H
H
5.5
±0.8)
bromides (15) in the R
3 5
and R positions were able to restore the
(
activity. However, by increasing the lipophilic bulk in the R
4
CH
3
N(CH
3
)
2
6.7
± 0.2)
position, Br (16) > Cl (17) = F (18) activity restored, indicative of
a critical hydrophobic binding interaction of the benzo-portion of
the quinoline scaffold. This was further supported by the
positioning of a methyl in the R positions (compound 19)
4
restored much of the activity. These findings indicated to us that
the binding pocket readily accepts larger hydrophobic moieties in
(
3
3
3
3
3
0
1
2
3
4
CH
H
3
H
H
H
H
H
CH(CH
3
)
2
H
H
H
H
H
7.8
±0.1)
(
C(CH
3
)
3
N(CH
N(CH
N(CH
N(CH
3
3
3
3
)
2
2
2
2
>25.0
>25.0
(CH
2
)
3
CH
3
H
)
)
)
3 5
the R -R area of the quinoline scaffold.
CH
2
CH
3
CH
2
CH
3
18.7
(±1.0)
Table 2. Structure and IC50 values of substituted quinolines for inhibition of
chymotrypic activity of the 20S proteasome. All IC50 values are averages of
two independent experiments (each performed in triplicate).
CH
3
10.0
±0.6)
(
3
3
5
6
H
H
N(CH
3
)
2
H
H
>25
CH
3
CH
3
CH
3
(CH
2
)
3
CH
3
21.6
±1.1)
(
R1
R2
-
R3
-
R4
-
R5
IC50
B T Z
-
0 . 0 0 6 2
-
37
-
-
-
H
H
CH -( CH
3
)
2
HX X
  X . X 
>25.0
>25.0

                                                                                 0 . 0
0
0 3 

3
8
Br
H
7
C H
3
C H
3
H
C H
3
    
     

8
9
CH
2
CH
3
CH
3
H
H
CH
3
19.9
±0.7)
(
This was further confirmed by the incorporation of a
hydrophobic butyl group in the R position, which exhibited
excellent activity (20, IC50 7.6 M). Similarly, the
dimethylamino moiety in the R position (22) successfully
encumbers the hydrophobic pockets occupied by the two methyl
groups in the R and R positions in compound 7. As seen
previously, minor changes in potency were seen following
changes in the R position (compounds 21-23). The incorporation
H
CH
3
CH
3
13.8
±1.3)
4
(
1
1
0
1
CH
2
CH
3
CH
3
3
H
H
CH
3
3
>25.0
4
CH
3
CH
CH
8.2
3
5
(±1.2)
1
2
CH
3
H
CH
3
>25.0
1
of piperidinyl group (25) further improved activity with an IC50
of 5.4 M. The replacement of the dimethylamino group by the
smaller methoxy moiety (26) eliminated all activity, which was
partially restored by additional neighboring methoxy-groups
1
1
3
4
CH
3
H
H
H
H
>25.0
H
Cl
Cl
>25.0
(27). Considering the lack of activity of 26, this restoration of
activity is likely due to positioning of the two methyl groups of
dimethylamine of compound 22 in two hydrophobic binding
pockets, rather than a possible hydrogen accepting role. This was
further confirmed with compound 30, which contains an
isopropyl moiety instead of the dimethylamino moiety and still
retains much of its activity (IC50 of 7.8 M). Considering the
1
1
5
6
H
Br
H
H
Br
H
>25.0
CH
3
Br
9.9
(± 0.6)
1
1
1
2
7
8
9
0
CH
CH
CH
CH
3
H
H
H
H
Cl
F
H
H
H
H
>25.0
3 5
apparent requirement of the alkyl groups in the R and R
positions in 7, it is likely that the two methyl groups on
dimethylamine 22 occupy similar pockets given the close special
proximity to the R and R position. As anticipated based on the
3 5
activity of fully saturated cyclohexyl (11), compounds 28-30 all
exhibit potent activities with IC50’s in the 5-7 M range. Further
1 2
functionalization around the R and R position (31-38), did not
3
3
3
>25.0
8.5
CH
3
(± 0.1)
(CH
2
)
3
CH
3
7.6
(± 1.3)
render more active analogues, although the trends of proteasome
activity followed our earlier findings.
2
1
H
H
N(CH
3
)
2
H
6.3
±0.3)
(

The lead compound, quinoline 25, was subsequently evaluated
for its inhibition of the proteasome’s tryptic (2)-like and caspase
(0.10 mmol) and dissolved in dry toluene (2 mL). The solution
was loaded with the aniline derivative (1.0 mmol), alkyne
derivative (1.0 mmol) and tert-butylisonitrile (1.5 mmol). The
pressure tube was sealed with a Teflon screw cap, taken out of
the dry box and heated at 100 °C for 24-48 h in a silicone oil
bath. Volatiles were removed in vacuo, and glacial acetic acid (2
mL) was added. The mixture was then heated at 150 °C for 24 h.
The pressure tube was then allowed to cool to room temperature,
diluted in dichloromethane and neutralized with saturated
(1)-like activity in vitro using purified human 20S proteasome.
Similar to quinoline 7 (Fig. 3A), compound 25 also inhibited
caspase-like activity (IC50 10.9 M), but not the tryptic-like
activity (suppl. Fig. S6).
3.
Conclusion
NaHCO
additional dichloromethane, washed with brine, dried over
NaSO , filtered and concentrated in vacuo. The crude product
was dry loaded onto alumina and purification was accomplished
by column chromatography on neutral alumina using
hexanes:ethyl acetate (9:1, v/v) as the eluent to provide the
desired quinoline.
3
solution. The organic layer was further extracted with
In conclusion, new classes of small molecule proteasome
inhibitors are direly needed considering the inevitable resistance
to current drugs and therapeutic limitations in the clinic. We
described herein a novel class of noncovalent nonpeptidic
proteasome inhibitors. The in vitro activity of our parent scaffold
translated well in cell culture, where quinoline 7 prevents NF-
B mediated gene expression in HeLa cells following TNF-
4
7

stimulation, consistent with a mechanism of proteasome
inhibition. Our SAR study identified several domains that were
found to be critical for the activity of this new lead template.
Quinoline 25 was found to be the most active analogue in this
series and inhibited the chymotryptic activity of the 20S
proteasome with an IC50 of 5.4 M. This potency is similar to the
4
.1.1. 5,7-dimethyl-3-phenylquinoline (4)
1
Pale yellow solid 128 mg (55%). H NMR (CDCl
C): δ = 2.51 (3 H, s, CH ), 2.68 (3 H, s, CH ), 7.23 (1 H, s, Ar-
H), 7.39-7.42 (1 H, m, Ar-H, 7.48-7.53 (2 H, m, Ar-H), 7.68-7.70
2 H, m, Ar-H), 7.74 (1 H, s, Ar-H), 8.36-8.37 (1H, d, J = 2 Hz,
3
, 500 MHz, 20
⁰
3
3
(
13
32
Ar-H), 9.10 (1 H, d, J = 2 Hz, Ar-H). C NMR (CDCl
MHz, 20 ⁰C): δ = 18.5, 21.8, 125.3, 126.4, 127.4, 127.8, 129.1
29.6, 129.8, 132.6, 134.2, 138.4, 139.3, 147.9, 149.3. MS (EI):
3
, 125
noncovalent proteasome inhibitors, PI-083 (IC50 ~1.0 M), 5-
37
AHQ (IC50 ~0.6-5 M), and a significant improvement over the
3
6
1
millimolar activity of chloroquine. Thus, these studies describe
a novel class of quinoline-based proteasome inhibitors. Further
optimization of this new template is currently under investigation
in our laboratories as well as the evaluation of the potential
clinical significance of this new nonpeptidic, noncovalent class
of proteasome inhibitors.
+
m/z 233 (M ). Anal. Found (Calcd): %C, 87.42 (87.52); %H, 6.52
(6.48); %N 6.06 (6.00). M.p.: 76-78 ⁰C.
4
.1.2. 2,6-dimethyl-3-phenylquinoline (5)
1
Pale yellow solid 158 mg (68%). H NMR (CDCl
C): δ = 2.50 (3 H, s, CH ), 2.63 (3 H, s, CH ), 7.37-7.39 (3 H,
m, Ar-H), 7.40-7.43 (2 H, m, Ar-H), 7.49-7.51 (2 H, m, Ar-H),
3
, 500 MHz, 20
⁰
3
3
4.
Experimental section
13
7.83 (1 H, s, Ar-H), 7.94-7.96 (1 H, d, J = 2 Hz, Ar-H). C NMR
All manipulations of air sensitive compounds were carried out in
an Mbraun drybox under a purified nitrogen atmosphere. Toluene
(CDCl , 125 MHz, 20 ⁰C): δ = 21.5, 24.4, 126.2, 126.7, 127.4,
3
128.0, 128.3, 129.1, 131.5, 135.4, 135.6, 135.7, 140.0, 145.6,
was purified by sparging with dry N
2
and water was removed by
+
156.2 MS (EI): m/z 233 (M ). Anal. Found (Calcd): %C, 87.62
running through activated alumina systems purchased from Solv-
1
13
19
(87.52); %H, 6.42 (6.47); %N 5.96 (6.00). M.p.: 80-81 ⁰C.
Tex. H, C and F spectra were recorded on VXR-500
spectrometers. Melting points are uncorrected and measured on a
Mel-Temp II apparatus (Laboratory Devices Inc, USA) with a
4.1.3. 2-ethyl-3-cyclohexenyl-5,7-dimethylquinoline (8)
1
Light yellow oil 102 mg (38%). H NMR (CDCl
C): δ = 1.51-1.61 (7 H, m) 1.99-2.03 (2 H, m, CH
(2 H, m, CH ), 2.21 (3 H, s, CH ), 2.31 (3 H, s, CH
2H, q, J = 8 Hz, CH ), 5.60-5.62 (1 H, m, CH), 6.86 (1 H, s, Ar-
H), 7.85 (1 H, s, Ar-H), 8.05 (1 H, s, Ar-H). C NMR (CDCl
125 MHz, 20 ⁰C): δ = 13.6, 18.0, 21.3, 22.0, 23.0, 25.3, 29.1,
3
, 500 MHz, 20
), 2.14-2.16
), 3.04-3.09
mercury thermometer in an open capillary tube. Ti(NMe
2 2
) dpma
⁰
2
and Ti(NMe dpm were made following the literature
2 2
)
4
4-46
2
3
3
procedures.
synthesized, with the exceptions 31 and 32 in which
Ti(NMe dpma was used. Tert-butylisonitrile was made
2 2
Ti(NMe ) dpm was used for all of the quinolines
(
2
1
3
3
,
2 2
)
54
according to the literature procedure and purified by distillation
under dry nitrogen but it may also be purchased from Sigma
Aldrich. Hexanes and ethyl acetate were purchased from
Mallinckrodt chemicals and used as received. Alkynes were
purchased either from Sigma Aldrich or from GFC chemicals and
were dried/distilled from CaO under dry nitrogen before use.
Amines were purchased from Sigma Aldrich, dried over KOH
and distilled under nitrogen. Palladium(II) acetate, potassium
tert-butoxide and 2-(dicyclohexylphosphino)biphenyl (97%)
were also purchased from Sigma Aldrich and used as received. 2-
methylquinoline (1), 3-methylquinoline (2), were purchased from
TCI America. 3-phenylquinoline (3) was synthesized through via
3
1
2
0.8, 124.3, 126.7, 126.8, 128.4, 130.5, 133.3, 136.5, 137.9,
38.0, 148.2, 160.9. MS (EI): m/z 265 (M ). HRMS, Found: m/z
65.1840.; Calcd for C19
+
+
H
23N 265.1830.
4
.1.4. 3-cyclohexenyl-5,7-dimethylquinoline (9)
1
Pale yellow solid 99 mg (42%). H NMR (CDCl
C): δ = 1.69-1.71 (2 H, m, CH ), 1.82-1.84 (2 H, m, CH
), 2.47 (3 H, s, CH ), 2.48-2.52 (2 H, m, CH
), 6.29-6.30 (1 H, m, CH), 7.17 (1 H, s, Ar-H),
.66 (1 H, s, Ar-H), 8.07-8.08 (1H, d, J = 2 Hz, Ar-H), 8.94 -8.95
3
, 500 MHz, 20
), 2.26-
),
⁰
2
2
2
2
7
.28 (2 H, m, CH
.63 (3 H, s, CH
2
3
2
3
13
(
1 H, d, J = 2 Hz, Ar-H). C NMR (CDCl
= 18.6, 21.7, 21.9, 22.9, 26.0, 27.3, 125.2, 126.1, 126.8, 126.9,
28.3, 129.5, 129.6, 133.9, 134.1, 134.3, 148.1. MS (EI): m/z 237
3
, 125 MHz, 20 ⁰C): δ
55
literature procedure. 3-cyclohexenyl-2,5,7-trimethylquinoline
7), 2,5,7-trimethyl-3-phenylquinoline (6), 5,7-dimethyl-2,3-
diphenylquinoline (12), 2-methyl-3-phenyl-6-(N,N-
dimethylamino)quinoline (34) were synthesized via literature
1
(
+
(M ). Anal. Found (Calcd): %C, 86.22 (86.03); %H, 7.69 (8.07);
%
N 5.82 (5.90). M.p.: 58-59 ⁰C.
43
procedure.
4
.1.5. 2-ethyl-5,7-dimethyl-3-(prop-1-en-2-yl)quinoline (10)
1
4.1 General procedure for the synthesis of quinolines (1-39)
Pale yellow solid 158 mg (62%). H NMR (CDCl
C): δ = 1.34-1.37 (3 H, t, J = 7.5 Hz, CH ), 2.12 (3 H, s, CH
2.46 (3H, s, CH ), 2.57 (3 H, s, CH ), 2.96-3.00 (2 H, q, J = 8 Hz,
3
, 500 MHz, 20
⁰
3
3
),
In a nitrogen filled glove box, a 10 mL pressure tube, equipped
with a magnetic stir bar was loaded with the titanium catalyst
3
3
CH ), 4.98-4.99 (1 H, d, J = 1 Hz, CH), 5.27-5.28 (1 H, d, J = 1
2

+
Hz, CH), 7.08 (1 H, s, Ar-H), 7.67 (1 H, s, Ar-H), 7.89 (1 H, s,
(EI): m/z 301 (M ).HRMS, Found: m/z 301.0480.; Calcd for
1
3
+
Ar-H). C NMR (CDCl
3
, 125 MHz, 20 ⁰C): δ = 14.1, 18.3, 21.6,
C
16
H
16BrN 301.0466.
2
1
4.9, 29.1, 116.1, 123.8, 125.7, 128.4, 130.7, 133.5, 135.7, 138.4,
44.5, 147.5, 160.6. MS (EI): m/z 255 (M ). Anal. Found
+
4.1.11. 6-chloro-2-methyl-3-cyclohexenylquinoline (17)
1
(Calcd): %C, 85.32 (85.2); %H, 8.53 (8.50); %N 6.15 (6.22).
Light yellow oil 54 mg (21%). H NMR (CDCl
C): δ = 1.72-1.75 (2 H, quin, J = 11 Hz, CH ), 1.79-1.82 (2 H,
quin, J = 12 Hz, CH ), 2.21 (2 H, m, CH ), 2.25 (2 H, m, CH ),
.67 (3 H, s, CH ), 6.32 (1 H, m CH), 7.54-7.57 (1 H, dd, J = 2
Hz, 9 Hz, Ar-H), 7.69 (1 H, s, Ar-H), 7.70-7.71 (1H, d, J = 3 Hz,
3
, 500 MHz, 20
⁰
2
2
2
2
4
3
.1.6.
3-cyclohexyl-2,5,7-trimethylquinoline (11)
2
3
-cyclohexenyl-2,5,7-trimethylquinoline (7) (60 mg, 0.024
mmol) was dissolved in 6 mL of ethanol and hydrogenated at low
pressure, using a hydrogen ballon, over 10% palladium on carbon
1
3
Ar-H), 7.91-7.93 (1 H, d, J = 9 Hz, Ar-H). C NMR (CDCl
MHz, 20 ⁰C): δ = 21.9, 22.9, 23.7, 25.4, 30.0, 125.8, 127.6,
27.9, 129.6, 129.8, 131.2, 133.5, 136.9, 139.1, 144.9, 158.2. MS
3
, 125
(
100 mg) at room temperature (25 ⁰C) for an hour. Purification
1
was accomplished via filtration through neutral alumina followed
+
(EI): m/z 257 (M ). HRMS, Found: m/z 257.0963.; Calcd for
by column chromatography on neutral alumina using
hexanes:ethyl acetate (9:1, v/v), which afforded the desired
compound as a pale white liquid 54 mg (90%). H NMR (CDCl
+
C
16
H
16ClN 257.0971.
1
3
,
4
.1.12. 3-cyclohexenyl-6-fluoro-2-methylquinoline (18)
5
00 MHz, 20 ⁰C): δ = 1.42-1.49 (4 H, m, CH
m, CH ), 1.89-1.95 (4 H, m, CH ), 2.46 (3 H, s, CH
s, CH ), 2.73 (3 H, s, CH ), 2.79-2.81 (1 H, m, CH), 7.10 (1 H, s,
Ar-H), 7.62 (1 H, s, Ar-H), 7.96 (1 H, s, Ar-H). C NMR
CDCl , 125 MHz, 20 ⁰C): δ = 18.5, 22.8, 26.2, 27.1, 29.7, 33.9,
0.1, 124.7, 125.4, 128.1, 128.4, 128.5, 133.5, 138.2, 143.3,
2
), 1.79-1.82 (2 H,
1
Brown oil 38 mg (15%). H NMR (CDCl
1.69-1.71 (2 H, m, CH ), 1.72-1.78 (2H, m, CH
H, m, CH ), 2.20-2.25 (2 H, m, CH ), 2.65 (3 H, s, CH ), 5.67-
3
, 500 MHz, 20 ⁰C): δ
2
2
3
), 2.61 (3 H,
=
2
2
), 2.17-2.19 (2
3
3
13
2
2
3
5.68 (1 H, m, CH), 7.29-7.31 (1 H, dd, J = 3 Hz, 9 Hz, Ar-H),
(
4
1
3
7
.34-7.38 (1 H, ddd, J = 3 Hz, 9 Hz, 9 Hz, Ar-H), 7.70 (1 H, s,
13
+
Ar-H), 7.94-7.97 (1 H, dd, J = 6 Hz, 10 Hz, Ar-H). C NMR
CDCl , 125 MHz, 20 ⁰C): δ = 21.9, 22.9, 23.5, 25.4, 30.0, 110.0-
10.1 (d, JCF = 21 Hz), 118.6-118.8 (d, JCF = 26 Hz), 127.4-127.5
d, JCF = 10 Hz), 127.7, 130.5-130.6 (d, JCF = 9 Hz), 133.7-133.8
(d, JCF = 5 Hz), 137.0, 138.9, 143.7, 157.0-157.1 (d d, J = 3
57.3. MS (EI): m/z 253 (M ). Anal. Found (Calcd): %C, 85.29
(
3
(85.32); %H, 9.23 (9.15); %N 5.48 (5.53).
1
(
4
.1.7.
Yellow liquid 60 mg (28%). H NMR (CDCl
δ = 1.68-1.72 (2 H, m, CH ), 1.76-1.80 (2 H, m, CH
), 2.23-2.26 (2 H, m, CH ), 2.67 (3 H, s, CH
.68 (1 H, m, CH), 7.40-7.43 (1 H, m, Ar-H), 7.58-7.61 (1H, m,
3-cyclohexenyl-2-methylquinoline (13)
1
CF
3
, 500 MHz, 20 ⁰C):
), 2.17-2.23
), 5.67-
19
Hz), 159.1. F NMR (CDCl
3
, 470 MHz, 20 ⁰C): δ = 115.0 (m).
MS (EI): m/z 241 (M ). Anal. Found (Calcd): %C, 79.59 (79.64);
H, 6.65 (6.68); %N 5.75 (5.80).
2
2
+
(2 H, m, CH
2
2
3
%
5
Ar-H), 7.69-7.71 (1 H, d, J = 9 Hz, Ar-H), 7.75 (1 H, s, Ar-H),
13
4.1.13. 3-cyclohexenyl-2,6-dimethylquinoline (19)
1
7
2
1
3
.96-7.98 (1 H, d, J = 9 Hz, Ar-H). C NMR (CDCl , 125 MHz,
0 ⁰C): δ = 21.9, 22.9, 23.7, 25.4, 30.1, 125.6, 126.9, 127.1,
Brown oil 123 mg (52%). H NMR (CDCl
1.68-1.72 (2 H, m, CH ), 1.75-1.78 (2 H, m, CH
H, m, CH ), 2.22-2.24 (2 H, m, CH ), 2.46 (3 H, s, CH ), 2.64 (3
3
, 500 MHz, 20 ⁰C): δ
=
2
2
), 2.17-2.20 (2
27.4, 128.2, 128.7, 134.3, 137.3, 138.1, 146.6, 157.6. MS (EI):
+
2
2
3
m/z 223 (M ). Anal. Found (Calcd): %C, 86.12 (86.06); %H, 7.60
H, s, CH
.66 (1 H, s, Ar-H), 7.85-7.86 (1 H, d, J = 9 Hz, Ar-H). C NMR
CDCl , 125 MHz, 20 ⁰C): δ = 21.4, 22.0, 22.9, 23.6, 25.4, 30.1,
126.0, 126.9, 127.3, 127.9, 130.9, 133.7, 135.3, 137.4, 138.1,
3
), 5.65-5.67 (1 H, m, CH), 7.42-7.45 (2 H, m, Ar-H),
(7.67); %N 6.28 (6.27).
13
7
(
3
4
.1.8.
Brown solid 131 mg (45%). H NMR (CDCl
δ = 1.70-1.73 (2 H, m, CH ), 1.77-1.80 (2 H, m, CH
), 2.23-2.26 (2 H, m, CH ), 2.65 (3 H, s, CH
.71 (1 H, m, CH), 7.46-7.47 (1 H, d, J = 2 Hz, Ar-H), 7.87-7.89
5,7-dichloro-3-cyclohexenyl-2-methylquinoline (14)
1
3
, 500 MHz, 20 ⁰C):
), 2.18-2.23
), 5.69-
+
1
8
45.3, 156.6. MS (EI): m/z 237 (M ). Anal. Found (Calcd): %C,
2
2
6.04 (86.03); %H, 8.09 (8.07); %N 5.87 (5.90).
(2 H, m, CH
2
2
3
5
13
4.1.14. 6-butyl-3-cyclohexenyl-2-methylquinoline (20)
1
3
(1H, m, J = 3 Hz, Ar-H), 8.06 (1H, s, Ar-H). C NMR (CDCl ,
Viscous yellow oil 87 mg (32%). H NMR (CDCl
3
, 500 MHz, 20
1
1
25 MHz, 20 ⁰C): δ = 21.9, 22.8, 23.6, 25.4, 29.9, 123.6, 126.4,
26.7, 128.3, 130.9, 131.6, 133.6, 136.7, 139.3, 147.2, 159.9.
⁰
C): δ = 0.94-0.96 (3 H, t, J = 7 Hz, CH ), 1.35-1.43 (2 H, sext, J
3
+
= 15 Hz, CH ), 1.65-1.71 (2 H, quin, J = 15 Hz, CH ), 1.71-1.76
2
2
MS (EI): m/z 292 (M ). Anal. Found (Calcd): %C, 65.81 (65.77);
H, 5.20 (5.17); %N 4.72 (4.79). M.p.: 56-58 ⁰C.
2
(2 H, quin, J = 11 Hz, CH ), 1.79-1.84 (2 H, quin, J = 10 Hz,
%
CH
.76-2.79 (2 H, t, J =8 Hz, CH
H, m, Ar-H), 7.73 (1 H, s, Ar-H) 7.91-7.93 (1 H, d, J = 9 Hz, Ar-
2 2 2 3
), 2.22 (2H, m, CH ), 2.27 (2 H, m, CH ), 2.68 (3 H, s, CH ),
2
2
), 5.70 (1 H, s, CH), 7.48-7.50 (2
4
.1.9.
Yellow solid 122 mg (32%). H NMR (CDCl
δ = 1.71-1.73 (2 H, m, CH ), 1.78-1.80 (2 H, m, CH
), 2.24-2.26 (2H, m, CH ), 2.66 (3 H, s, CH
.72 (1 H, m, CH), 7.80-7.81 (1 H, d, J = 2 Hz, Ar-H), 8.01 (1 H,
5,7-dibromo-3-cyclohexenyl-2-methylquinoline (15)
1
3
, 500 MHz, 20 ⁰C):
), 2.20-2.23
), 5.71-
1
3
H). C NMR (CDCl
3
, 125 MHz, 20 ⁰C): δ = 14.0, 22.1, 22.3,
3.0, 23.6, 25.5, 30.2, 33.5, 35.6, 125.5, 126.9, 127.3, 127.9,
30.4, 134.0, 137.4, 138.1, 140.3, 145.4, 156.7. MS (EI): m/z 279
2
2
2
1
(2 H, m, CH
2
2
3
5
+
+
13
20 26
(M ). HRMS, Found: m/z 280.2062; Calcd for C H N
3
s, Ar-H), 8.11-8.12 (1 H, d, J = 2 Hz, Ar-H). C NMR (CDCl ,
280.2065.
1
1
25 MHz, 20 ⁰C): δ = 21.9, 22.8, 23.5, 25.4, 29.9, 121.8, 121.9,
25.2, 128.4, 130.7, 132.1, 133.5, 136.7, 139.8, 147.4, 159.9. MS
+
4.1.15. 3-cyclohexenyl-6-(N,N-dimethylamino)quinoline (21)
(EI): m/z 381 (M ). Anal. Found (Calcd): %C, 50.51 (50.43);
1
Viscous yellow oil 97 mg (38%). H NMR (CDCl
C): δ = 1.41-1.46 (2 H, m, CH ), 1.52-1.57 (2 H, m, CH
.97 (2 H, m, CH ), 2.23-2.4 (2 H, m, CH ), 2.51 (6 H, s,
N(CH ) ), 6.08-6.1 (1 H, m, CH), 6.64-6.65 (1 H, d, J = 3 Hz, Ar-
3
, 500 MHz, 20
%H, 3.93 (3.97); %N 3.64 (3.68). M.p.: 89-90 ⁰C.
⁰
1
2
2
), 1.95-
2
2
4
.1.10. 6-bromo-3-cyclohexenyl-2-methylquinoline (16)
1
3
2
Light brown oil 103 mg (34%) H NMR (C
C): δ = 1.45-1.54 (4 H, m, CH ), 1.93-1.96 (4 H, m, CH
3 H, s, CH ), 5.41-5.42 (1 H, m, CH), 7.13 (1 H, s, Ar-H), 7.38-
.40 (1 H, dd, J = 2 Hz, 9 Hz, Ar-H), 7.58-7.59 (1 H, d, J = 2 Hz,
6
D
6
, 500 MHz, 20
H), 6.95-6.97 (1 H, dd, J = 3 Hz, 9 Hz, Ar-H), 7.67-7.69 (1 H, d,
J = 2 Hz, Ar-H), 8.26-8.28 (1 H, d, J = 9 Hz, Ar-H), 9.03-9.04 (1
⁰
(
7
2
2
), 2.60
3
1
3
H, d, J = 2 Hz, Ar-H). C NMR (CDCl
3
, 125 MHz, 20 ⁰C): δ =
1
3
22.0, 22.8, 25.8, 26.9, 39.9, 105.3, 118.5, 125.9, 128.2, 129.6,
Ar-H), 7.88-7.90 (1 H, d, J = 9 Hz, Ar-H). C NMR (C
MHz, 20 ⁰C): δ = 21.9, 22.8, 23.6, 25.3, 29.7, 119.2, 127.2,
28.2, 129.2, 130.7, 131.8, 132.7, 137.1, 138.6, 145.7, 157.8. MS
6 6
D , 125
+
1
30.1, 134.3, 135.0, 142.1, 144.9, 148.6. MS (EI): m/z 252 (M ).
+
20 2
HRMS, Found: m/z 252.1634.; Calcd for C17H N 252.1626.
1

1
the desired compound as a viscous brown oil 42 mg (42%). H
NMR (CDCl , 500 MHz, 20 ⁰C): δ = 1.56-1.60 (2 H, m, CH ),
1.61-1.78 (8 H, m, CH ), 2.17-2.23 (4 H, m, CH ), 2.61 (3 H, s,
CH ), 3.21-3.23 (4 H, m, NCH ), 5.64-5.65 (1 H, m, CH), 6.94-
6.95 (1 H, d, J = 3 Hz, Ar-H), 7.51-7.54 (1 H, m, Ar-H), 7.60 (1
4
.1.16. 3-cyclohexenyl-2-methyl-6-(N,N-dimethylamino)
quinoline (22)
Brown solid 117 mg (44%). H NMR (CDCl
δ = 1.72-1.74 (2 H, m, CH ), 1.75-1.78 (2H, m, CH
), 2.25-2.29 (2H, m, CH ), 2.64 (3 H, s, CH
6 H, s, N(CH ), 5.68-5.70 (1 H, m, CH), 6.78-6.79 (1 H, d, J =
Hz, Ar-H), 7.29-7.32 (1 H, dd, J = 3 Hz, 9 Hz, Ar-H), 7.63 (1
3
2
2
2
1
3
, 500 MHz, 20 ⁰C):
), 2.22-2.24
), 3.04
3
2
2
2
13
(2 H, m, CH
2
2
3
H, s, Ar-H), 7.82-7.84 (2 H, d, J = 10 Hz, Ar-H). C NMR
(
3
)
3 2
(CDCl , 125 MHz, 20 ⁰C): δ = 22.0, 23.0, 23.2, 24.3, 25.4, 25.7,
3
30.1, 50.8, 128.3, 128.6, 129.6, 130.0, 132.9, 133.3, 137.6, 138.2,
13
+
H, s, Ar-H), 7.86-7.88 (1 H, d, J = 9 Hz, Ar-H). C NMR
CDCl , 125 MHz, 20 ⁰C): δ = 22.1, 23.0, 23.2, 25.4, 30.1, 40.8,
05.2, 118.8, 126.8, 128.2, 128.8, 132.7, 137.7, 138.3, 140.8,
149.7, 154.2, 166.5. MS (EI): m/z 306 (M ). Anal. Found
(
3
(Calcd): %C, 82.36 (82.31); %H, 8.49 (8.55); %N 9.15 (9.14).
1
1
8
+
48.2, 153.1. MS (EI): m/z 266 (M ). Anal. Found (Calcd): %C,
1.14 (81.16); %H, 8.40 (8.32); %N 10.46 (10.52). M.p.: 81-83
4.1.20. 3-cyclohexenyl-6-methoxy-2-methylquinoline (26)
1
Brown solid 108 mg (43%). H NMR (CDCl , 500 MHz, 20 ⁰C):
3
⁰
C.
δ = 1.72-1.75 (2 H, m, CH
2 H, m, CH ), 2.25-2.28 (2 H, m, CH
(3H, s, OCH ), 5.69-5.70 (1 H, m, CH), 7.00-7.01 (1 H, d, J = 3
Hz, Ar-H), 7.27-7.30 (1 H, dd, J = 3 Hz, 9 Hz, Ar-H), 7.69 (1 H,
s, Ar-H), 7.89-7.91 (1 H, d, J = 9 Hz, Ar-H). C NMR (CDCl ,
3
125 MHz, 20 ⁰C): δ = 22.0, 22.9, 23.3, 25.4, 30.1, 55.4, 104.8,
2
), 1.80-1.82 (2 H, m, CH
2
), 2.22-2.23
(
2
2
), 2.66 (3 H, s, CH
3
), 3.89
4
.1.17. 3-cyclohexenyl-2-ethyl-6(N,N-dimethylamino)
quinoline (23)
Viscous light yellow oil 109 mg (39%). H NMR (CDCl
MHz, 20 ⁰C): δ = 1.52-1.60 (7 H, m), 1.98-2.01 (2 H, m, CH
3
1
13
3
, 500
),
), 3.03-3.07 (2 H,
), 5.59 (1 H, m), 6.65 (1 H, d, J = 3 Hz, Ar-
2
2
.12-2.17 (2 H, m, CH
2
), 2.52 (6 H, s, N(CH
)
3 2
121.1, 127.2, 127.7, 129.7, 133.4, 137.4, 138.4, 142.7, 154.9,
+
quart, J = 7 Hz, CH
2
157.1. MS (EI): m/z 253 (M ). Anal. Found (Calcd): %C, 80.49
H), 6.98-7.00 (1 H, dd, J = 3 Hz, 9 Hz, Ar-H), 7.50 (1 H, s, Ar-
(80.60); %H, 7.60 (7.56); %N 5.57 (5.53). M.p.: 50-52 ⁰C.
1
3
H), 8.20-8.22 (1 H, d, J = 9 Hz, Ar-H). C NMR (CDCl
3
, 125
MHz, 20 ⁰C): δ = 13.7, 22.1, 23.1, 25.4, 28.9, 30.7, 40.1, 105.1,
4.1.21. 3-cyclohexenyl-5,6,7-trimethoxy-2-methylquinoline
1
1
18.7, 126.3, 128.2, 128.4, 129.7, 132.7, 138.0, 141.9, 148.0,
(27)
+
1
57.2. MS (EI): m/z 280 (M ). HRMS, Found: m/z 280.1928.;
Viscous light yellow oil 131 mg (42%). H NMR (CDCl
MHz, 20 ⁰C): δ = 1.71-1.76 (2 H, m, CH ), 1.79-1.84 (2 H, m,
3
, 500
+
2
Calcd for C19
H
24
N
280.1939.
2
CH
3.97 (3 H, s, OCH
5.69 (1 H, s, Ar-H), 7.20 (1 H, s, Ar-H), 7.98 (1 H, s, Ar-H).
2
), 2.23 (2 H, m, CH
2
), 2.27 (2 H, m, CH
2
), 2.65 (3 H, s, CH
3
),
),
C
4
(
.1.18. 3-cyclohexenyl-2-methyl-6-morpholinylquinoline
24)
A pressure tube was loaded with Pd(OAc)
3
), 3.99 (3 H, s, OCH
3
), 4.06 (3 H, s, OCH
3
1
3
2
(0.4 mg, 2 nmol), 2-
NMR (CDCl , 125 MHz, 20 ⁰C): δ = 22.0, 23.0, 23.4, 25.5, 30.3,
3
t
(dicyclohexylphosphino)biphenyl (1.4mg, 4 nmol), and KO Bu
56.1, 61.2, 61.6, 103.2, 117.8, 127.4, 129.0, 136.0, 137.5, 140.3,
+
(53 mg, 48 mmol) under nitrogen atmosphere. Anhydrous
144.3, 146.8, 155.4, 156.8. MS (EI): m/z 313 (M ). HRMS,
+
3
toluene was added followed by 6-bromo-3-cyclohexenyl-2-
methylquinoline (120 mg, 40 mmol) and morpholine (41 μL, 48
mmol). The tube was sealed, and then the mixture was stirred for
Found: m/z 314.1763.; Calcd for C H NO 314.1756. M.p.: 93-
1
9
24
95 ⁰C.
1
8 h at 110 ⁰C. After cooling,, the mixture was diluted with
dichloromethane (20 mL) and washed with water (20 mL), and
then brine (20 mL). The organic phase was dried over MgSO
4.1.22. 3-cyclohexyl-6-(N,N-dimethylamino)quinoline (28)
3-cyclohexenyl-6-(N,N-dimethylamino)quinoline (21) (80 mg,
0.32 mmol) was dissolved in 6 mL of ethanol and hydrogenated
at low pressure, using a hydrogen ballon, over 10% palladium on
carbon (120 mg) at room temperature (25 ⁰C) overnight.
Purification was accomplished via filtration through neutral
alumina followed by column chromatography on neutral alumina
using hexanes:ethyl acetate (9:1, v/v), which afforded the desired
4
,
and then the solvent was removed in vacuo. Purification was
accomplished by column chromatography on neutral alumina.
The eluent was hexanes:ethyl acetate (19:1, v/v), which afforded
1
the desired compound as a viscous brown oil 63 mg (51%). H
NMR (CDCl
.18-2.22 (4 H, m, CH
NCH ), 3.85-3.89 (4 H, m, OCH2), 5.63-5.67 (1 H, m, CH), 6.94-
.95 (1 H, d, J = 3 Hz, Ar-H), 7.63 (1 H, s, Ar-H), 7.88-7.90 (2
3
, 500 MHz, 20 ⁰C): δ = 1.68-1.78 (4 H, m, CH
2
),
1
2
2
), 2.62 (3 H, s, CH ), 3.21-3.23 (4 H, m,
3
compound as a viscous light yellow oil 75 mg (93%). H NMR
2
(CDCl , 500 MHz, 20 ⁰C): δ = 1.07-1.11 (1 H, m, CH ), 1.18-
3
2
6
2 2
1.22 (2 H, m, CH ), 1.27-1.33 (2 H, m, CH ), 1.58-1.60 (1 H, m,
CH ), 1.64-1.68 (2 H, m, CH ), 1.74-1.77 (2 H, m, CH ), 2.33-
2 2 2
2.38 (1 H, m, CH), 2.52 (6 H, s, N(CH ) ), 6.65-6.66 (1 H, d, J =
3 2
1
3
H, d, J = 5 Hz, Ar-H), 8.00-8.02 (2H, d, J = 5 Hz, Ar-H).
NMR (CDCl , 125 MHz, 20 ⁰C): δ = 22.0, 23.1, 25.4, 29.6, 30.1,
9.6, 66.8, 121.5, 127.2, 127.8, 128.2, 128.8, 129.6, 132.9, 133.5,
C
3
4
1
3 Hz, Ar-H), 6.97-6.99 (1 H, dd, J = 3 Hz, 9 Hz, Ar-H), 7.52 (1
H, d, J = 2 Hz, Ar-H), 8.26-8.28 (1 H, d, J = 9 Hz, Ar-H), 8.71-
+
38.5, 148.8, 154.7. MS (EI): m/z 308 (M ). Anal. Found
13
(Calcd): %C, 77.81 (77.89); %H, 7.80 (7.84); %N 9.01 (9.08).
3
8.72 (1 H, d, J = 2 Hz, Ar-H). C NMR (CDCl , 125 MHz, 20
⁰
C): δ = 26.0, 26.7, 34.0, 40.0, 42.0, 105.1, 118.4, 129.8, 128.9,
+
4
.1.19. 3-cyclohexenyl-2-methyl-6-piperidinylquinoline (25)
130.2, 140.2, 142.1, 147.4, 148.4. MS (EI): m/z 254 (M ).
HRMS, Found: m/z 254.1774.; Calcd for C H N 254.1783.
+
2
A pressure tube was loaded with Pd(OAc) (0.3 mg, 1.6 nmol), 2-
2
1
7
22
t
(
(
dicyclohexylphosphino)biphenyl (1.1mg, 3.3 nmol), and KO Bu
44 mg, 39 mmol) under nitrogen atmosphere. Anhydrous
4.1.23. 3-cyclohexyl-2-methyl-6-(N,N-
dimethylamino)quinoline (29)
toluene was added followed by 6-bromo-3-cyclohexenyl-2-
methylquinoline (100 mg, 33 mmol) and piperidine (40 μL, 40
mmol). The tube was sealed, and then the mixture was stirred for
2-methyl-3-cyclohexenyl-6-(N,N-dimethylamino)quinoline (22)
(100 mg, 0.38 mmol) was dissolved in 6 mL of ethanol and
hydrogenated at low pressure, using a hydrogen ballon, over 10%
palladium on carbon (150 mg) at room temperature (25 ⁰C)
overnight. Purification was accomplished via filtration through
neutral alumina followed by column chromatography on neutral
alumina using hexanes:ethyl acetate (9:1, v/v), which afforded
1
8 h at 110 ⁰C. After cooling,, the mixture was diluted with
dichloromethane (20 mL) and washed with water (20 mL), and
then brine (20 mL). The organic phase was dried over MgSO
4
,
and then the solvent was removed in vacuo. Purification was
accomplished by column chromatography on neutral alumina.
The eluent was hexanes:ethyl acetate (19:1, v/v), which afforded
1
the desired compound as a viscous yellow oil 98 mg (97%). H

+
NMR (CDCl
3
, 500 MHz, 20 ⁰C): δ = 1.16-1.33 (4 H, m, CH
2
),
),
m/z 324 (M ). HRMS, Found: m/z 324.1642.; Calcd for
+
1
6
9
.63-1.79 (4 H, m, CH ), 2.55 (6 H, s, CH ), 2.71 (3 H, s, CH
2
3
3
C
23 20
H N
2
324.1626. M.p.: 159-161 ⁰C.
.72-6.73 (1 H, d, J = 3 Hz, Ar-H), 6.99-7.01 (1 H, dd, J = 3 Hz,
Hz, Ar-H), 7.63 (1 H, s, Ar-H), 8.20-8.22 (1 H, d, J = 9 Hz, Ar-
4.1.29. 6-butyl-2-methyl-3-phenylquinoline (36)
Viscous yellow oil 101 mg (37%). H NMR (CDCl
13
1
H). C NMR (CDCl
3
, 125 MHz, 20 ⁰C): δ = 25.6, 26.3, 27.0,
3
, 500 MHz,
3
1
2
3.7, 39.9, 40.2, 105.3, 118.4, 129.0, 129.5, 129.6, 139.2, 141.3,
20 ⁰C): δ = 0.95-0.98 (3H, t, J =7 Hz, CH
J = 10 Hz, CH ), 1.68-1.74 (2 J, quin, J = 10 Hz, CH
s, CH ), 2.79-2.82 (2 H, t, J = 5 Hz, CH ), 7.41-7.44 (3 H, m, Ar-
H), 7.47-7.50 (2 H, m, Ar-H), 7.56-7.57 (2 H, m, Ar-H), 7.91 (1
3
), 1.37-1.45 (2 H, sext,
+
48.0, 153.4. MS (EI): m/z 268 (M ). HRMS, Found: m/z
2
2
), 2.67 (3 H,
+
68.1934.; Calcd for C18
H
24
N
2
268.1939.
3
2
13
4
.1.24. 3-cyclohexenyl-6-isopropyl-2-methylquinoline (30)
H, s, Ar-H), 7.99-8.01 (1 H, d, J = 9 Hz, Ar-H). C NMR
1
Viscous yellow oil 112 mg (43%). H NMR (CDCl
0 ⁰C): δ = 1.32-1.34 (6 H, d, J = 7 Hz, CH ), 1.71-1.75 (2 H,
quin, J =11 Hz, CH ), 1.78-1.83 (2 H, quin, J = 12 Hz, CH ),
), 2.26 (2 H, m, CH ), 2.68 (3 H, s, CH ), 3.02-
.11 (1 H, sept, J = 7 Hz, CH), 5.69 (1 H, s, CH), 7.53 (1 H, s,
3
, 500 MHz,
(CDCl , 125 MHz, 20 ⁰C): δ = 14.0, 22.4, 24.4, 33.5, 35.6, 125.7,
3
2
3
126.8, 127.5, 128.1, 128.4, 129.2, 131.0, 135.6, 135.7, 140.1,
+
2
2
140.8, 145.8, 156.3. MS (EI): m/z 275 (M ). HRMS, Found: m/z
+
2
3
.21 (2 H, m, CH
2
2
3
276.1750.; Calcd for C20
H
22N 276.1752.
Ar-H), 7.53-7.55 (1H, app d, J = 9 Hz, Ar-H), 7.75 (1 H, s, Ar-
4.1.30. 6-isopropyl-2-methyl-3-phenylquinoline (37)
1
13
H), 7.94-7.96 (1 H, d, J = 9 Hz, Ar-H). C NMR (C
6
D
6
, 125
Viscous yellow oil 143 mg (55%). H NMR (CDCl
3
, 500 MHz,
),
MHz, 20 ⁰C): δ = 22.0, 23.0, 23.5, 23.7, 25.3, 30.0, 34.0, 123.3,
20 ⁰C): δ = 1.35-1.37 (2 H, d, J = 7 Hz, CH ), 2.67 (3 H, s, CH
3
3
1
1
26.8, 127.1, 128.3, 129.0, 133.6, 137.7, 137.8, 145.7, 146.5,
3.06-3.15 (1 H, sept, J = 8 Hz, CH), 7.40-7.44 (3 H, m, Ar-H),
7.47-7.50 (2 H, m, Ar-H), 7.60 (1 H, s, Ar-H), 7.61-7.64 (1 H,
dd, J = 9 Hz, Ar-H), 7.93 (1 H, s, Ar-H), 8.01-8.03 (1 H, d, J = 9
+
56.3. MS (EI): m/z 265 (M ). HRMS, Found: m/z 266.1907.;
+
2
Calcd for C15
H
20
N
266.1909.
13
Hz, Ar-H). C NMR (CDCl , 125 MHz, 20 ⁰C): δ = 23.9, 24.5,
3
4
.1.25. 3-tert-butyl-6-(N,N-dimethylamino)quinoline (31)
29.7, 34.1, 123.5, 126.8, 127.5, 128.2, 128.4, 129.2, 129.3, 135.6,
1
+
Viscous light brown oil 55 mg (24%). H NMR (CDCl
MHz, 20 ⁰C): δ = 1.20 (9 H, s, C(CH ), 2.53 (6H, s, N(CH
.67-6.68 (1 H, d, J = 3 Hz, Ar-H), 6.98-7.01 (1 H, dd, J = 3
Hz, 9 Hz, Ar-H), 7.72-7.73 (1 H, d, J = 3 Hz, Ar-H), 8.26-
.27 (1 H, d, J = 9 Hz, Ar-H), 8.96-8.98 (1 H, d, J = 2 Hz, Ar-
3
, 500
135.9, 140.1, 145.9, 146.7, 156.4. MS (EI): m/z 261 (M ).
+
)
3 3
3
)
2
),
HRMS, Found: m/z 262.1596.; Calcd for C19
H
20N 262.1593.
6
4.1.31. 6-bromo-2-methyl-3-phenylquinoline (38)
Light tan solid 337 mg (26%, on 5 mmol scale). H NMR
CDCl , 500 MHz, 20 ⁰C): δ = 2.66 (3 H, s, CH ), 7.39-7.41 (2H,
m, Ar-H), 7.43-7.46 (1 H, m, Ar-H), 7.48-7.52 (2 H, m, Ar-H),
1
8
13
(
3
3
H). C NMR (CDCl
3
, 125 MHz, 20 ⁰C): δ = 30.6, 33.3, 40.0,
105.3, 118.4, 128.4, 129.5, 130.0, 141.5, 143.0, 146.0, 148.5. MS
+
7
7
.76-7.78 (1 H, dd, J = 2 Hz, 9 Hz, Ar-H), 7.87 (1 H, s, Ar-H),
.93-7.95 (1 H, d, J = 9 Hz, Ar-H), 7.95-7.96 (1H, d, J = 2 Hz,
(EI): m/z 228 (M ). HRMS, Found: m/z 228.1617.; Calcd for
+
2
C
15 20
H N
228.1626.
1
3
Ar-H). C NMR (CDCl
3
, 125 MHz, 20 ⁰C): δ = 24.7, 119.7,
1
1
2
27.8, 128.0, 128.5, 129.1, 129.4, 130.2, 132.7, 134.9, 136.6,
4
.1.26. 2-butyl-6-(N,N-dimethylamino)quinoline (32)
+
1
39.4, 145.6, 158.0. MS (EI): m/z 297 (M ). HRMS, Found: m/z
Viscous brown oil 69 mg (30%). H NMR (CDCl
C): δ = 0.86-0.89 (3 H, t, J = 7 Hz, CH ), 1.32-1.39 (2 H, sext, J
7 Hz, CH ), 1.82-1.88 (2H, quin, J = 7 Hz, CH ), 2.50 (6 H, s,
N(CH ), 2.92-2.95 (2 H, t, J = 7 Hz, CH ), 6.64-6.65 (1 H, d, J
3 Hz, Ar-H), 6.92-6.93 (1 H, d, J = 8 Hz, Ar-H), 6.98-7.01 (1
H, dd, J = 3 Hz, 9 Hz, Ar-H), 7.61-7.63 (1 H, d, J = 8 Hz, Ar-H),
3
, 500 MHz, 20
+
98.0230.; Calcd for C16H13BrN 298.0231. M.p.: 88-90 ⁰C.
⁰
=
3
2
2
4
4
.2 Biological assays
)
3 2
2
=
.2.1 20S Proteasomal activity measurement
13
The fluorogenic substrates Suc-LLVY-AMC, Z-ARR-AMC, and
Z-LLE-AMC were used to measure CT-L, T-L and casp-L
proteasome activities, respectively. Assays were carried out in
black, clear bottom 96 well plates in a 200 L reaction volume
containing 1 nM of purified human 20S proteasome in 50 mM
Tris-HCL pH 7.5 and 0.03% SDS containing 50 M fluorogenic
substrate at 37° C. The rate of cleavage of fluorogenic peptide
substrates was determined by monitoring the fluorescence of
8
2
1
3
.19-8.21 (1 H, d, J = 9 Hz, Ar-H). C NMR (CDCl , 125 MHz,
0 ⁰C): δ = 13.9, 22.6, 31.9, 39.5, 40.0, 105.4, 119.1, 121.4,
28.1, 130.1, 133.8, 142.9, 148.0, 158.4. MS (EI): m/z 228 (M ).
+
+
2
HRMS, Found: m/z 228.1616.; Calcd for C15
H
20
N
228.1626.
4.1.27. 2,3-diethyl-N,N-dimethylquinoline (33)
1
Light brown solid 74 mg (33%). H NMR (CDCl
C): δ = 1.32-1.35 (3 H, t, J = 7 Hz, CH
Hz, CH ), 2.78-2.83 (2 H, q, J = 7 Hz, CH
7 Hz, CH ), 3.06 (6 H, s, N(CH ), 6.79 (1 H, d, J = 2.8 Hz,
Ar-H), 7.29-7.31 (1 H, dd, J = 3 Hz, 9 Hz, Ar-H), 7.71 (1 H, s,
3
, 500 MHz, 20
⁰
3
), 1.35-1.38 (3 H, t, J = 7
), 2.95-2.99 (2 H, q, J
released aminomethylcoumarin using
a SpectraMax M5e
3
2
multiwall plate reader at an excitation wavelength of 380 nm and
emission wavelength of 460 nm. Fluorescence was measured
every minute over a period of 30 minutes and the maximum
increase in fluorescence per minute was used to calculate specific
activities of each sample.
=
2
3 2
)
1
3
Ar-H), 7.89-7.91 (1 H, d, J = 9 Hz, Ar-H). C NMR (CDCl
MHz, 20 ⁰C): δ = 13.9, 14.5, 25.1, 28.5, 41.9, 105.0, 118.6,
3
, 125
+
128.7, 128.9, 132.5, 135.2, 148.2, 158.8. MS (EI): m/z 228 (M ).
+
2
HRMS, Found: m/z 229.1712.; Calcd for C15
M.p.: 64-66 ⁰C.
H
20
N
229.1705.
4
.2.2
HeLa NF-B-luc cells (5.0 x 10 cells/mL) were seeded into a
6-well white opaque plate using DMEM medium supplemented
NF-B-luc Reporter Assay
5
9
4.1.28. 2,3-diphenyl-6-(N,N-dimethylamino)quinoline (35)
1
with 5% fetal bovine serum, 100 U/mL penicillin, 100 g/mL
streptomycin, 1 mM sodium pyruvate, 0.2 mM L-glutamine, and
Dark yellow solid 90 mg (28%). H NMR (CDCl
C): δ = 2.52 (1 H, s, N(CH ), 6.63-6.64 (1 H, d, J = 3 Hz, Ar-
H), 7.01-7.06 (5 H, m, Ar-H), 7.07-7.10 (2 H, m, Ar-H), 7.19-
.21 (2 H, m, Ar-H), 7.72-7.74 (3 H, m, Ar-H), 8.30-8.32 (1 H, d,
3
, 500 MHz, 20
⁰
3 2
)
1
00 g /mL hygromycin B. After 24 hours the cell culture
medium was replaced with DMEM medium supplemented with
00 U/mL penicillin and 100 g/mL streptomycin. Cell cultures
7
1
3
1
J = 9 Hz, Ar-H). C NMR (CDCl
3
, 125 MHz, 20 ⁰C): δ = 39.9,
04.6, 119.4, 126.7, 127.2, 127.53, 128.0, 128.9, 129.8, 130.4,
30.5, 134.7, 135.6, 141.2, 141.4, 142.3, 148.5, 153.9. MS (EI):
1
1
were pretreated with vehicle (1% DMSO), 50 M peptide
aldehyde 12 (positive control) or quinoline (final concentrations
were 50, 25, 12.5, 6.25, 3.13 and 1.56 M) for 30 minutes at 37

ºC in 5% CO . TNF- was added to a final concentration of 25
(21)
Radhakrishnan, S. K.; Lee, C. S.; Young, P.; Beskow, A.;
2
ng/mL and the samples were further incubated for 8 hours at 37
Chan, J. Y.; Deshaies, R. J. Mol Cell 2010, 38, 17.
(22)
ºC in 5% CO . Cells were assayed for firefly luciferase
2
Schwartz, R.; Davidson, T. Oncology (Williston Park)
production using the Steady-Glo luciferase reporter assay
2004, 18, 14.
(
Promega, Madison, WI) according to manufacturer’s protocol.
(
(
23)
24)
Groll, M.; Huber, R.; Moroder, L. J Pept Sci 2009, 15, 58.
Gallastegui, N.; Beck, P.; Arciniega, M.; Huber, R.;
The luminescence of each well was measured using a Veritas
microplate luminometer. All reported data are the average of two
independent experiments unless otherwise indicated. The data
was analyzed using GraphPad Prism 4.0. The data was
normalized to TNF- activation and the EC values were
Hillebrand, S.; Groll, M. Angew Chem Int Ed Engl 2012, 51, 247.
(25) Beck, P.; Lansdell, T. A.; Hewlett, N. M.; Tepe, J. J.;
Groll, M. Angew Chem Int Ed Engl 2015.
(26) Ozcan, S.; Kazi, A.; Marsilio, F.; Fang, B.; Guida, W. C.;
Koomen, J.; Lawrence, H. R.; Sebti, S. M. J Med Chem 2013, 56,
783.
27)
Weilbaecher, K. N.; Henry, R. W.; Tepe, J. J. ACS Chem Biol 2013,
, 578.
28)
5
0
calculated using the equation for the sigmodial curve for variable
slope.
3
(
Supplemental data
Lansdell, T. A.; Hurchla, M. A.; Xiang, J.; Hovde, S.;
Supplemental data associated with this article include the IC50
inhibition curves and error analysis is available online at:
8
(
Azevedo, L.; Lansdell, T.; Ludwig, J.; Woloch, D.; Cogan,
Acknowledgments
D.; Patten, G.; Kuszpit, M.; Fisk, J.; Mosey, R. A.; Tepe, J. J. J. Med.
Chem. 2013, 56, 5974.
Financial support for this work was provided in part by the
National Science Foundation CHE-1265738 (ALO) and National
Institutes of Health CA-142644-01 (JJT).
(
29)
Basse, N.; Montes, M.; Marechal, X.; Qin, L.; Bouvier-
Durand, M.; Genin, E.; Vidal, J.; Villoutreix, B. O.; Reboud-Ravaux,
M. J Med Chem 2010, 53, 509.
(
30)
Hasegawa, M.; Yasuda, Y.; Tanaka, M.; Nakata, K.;
References
Umeda, E.; Wang, Y.; Watanabe, C.; Uetake, S.; Kunoh, T.;
Shionyu, M.; Sasaki, R.; Shiina, I.; Mizukami, T. Eur J Med Chem
(
1)
Murata, S.; Yashiroda, H.; Tanaka, K. Nat. Rev. Mol. Cell
2014, 71, 290.
Biol. 2009, 10, 104.
(
3
(
(
(
31) Ge, Y.; Kazi, A.; Marsilio, F.; Luo, Y.; Jain, S.; Brooks,
2)
3.
3)
4)
Groll, M.; Huber, R. Biochim. Biophys. Acta 2004, 1695,
W.; Daniel, K. G.; Guida, W. C.; Sebti, S. M.; Lawrence, H. R. J.
Med. Chem. 2012, 55, 1978.
Ben-Nissan, G.; Sharon, M. Biomolecules 2014, 4, 862.
Groll, M.; Heinemeyer, W.; Jager, S.; Ullrich, T.; Bochtler,
(
32)
Lawrence, H. R.; Kazi, A.; Luo, Y.; Kendig, R.; Ge, Y.;
Jain, S.; Daniel, K.; Santiago, D.; Guida, W. C.; Sebti, S. M. Bioorg.
Med. Chem. 2010, 18, 5576.
M.; Wolf, D. H.; Huber, R. Proc. Natl. Acad. Sci. U S A 1999, 96,
0976.
5)
Bartunik, H. D.; Huber, R. Nature 1997, 386, 463.
1
(
(
33)
Springett, G. M.; Berndt, N.; Yip, R. M.; Sebti, S. M. Cell Cycle
009, 8, 1940.
34) Beck, P.; Reboud-Ravaux, M.; Groll, M. Angew Chem Int
Ed Engl 2015, 54, 11275.
35) Gaczynska, M.; Osmulski, P. A. Antioxid Redox Signal
014, 21, 2286.
36) Sprangers, R.; Li, X.; Mao, X.; Rubinstein, J. L.;
Schimmer, A. D.; Kay, L. E. Biochemistry 2008, 47, 6727.
37) Li, X.; Wood, T. E.; Sprangers, R.; Jansen, G.; Franke, N.
Kazi, A.; Lawrence, H.; Guida, W. C.; McLaughlin, M. L.;
Groll, M.; Ditzel, L.; Lowe, J.; Stock, D.; Bochtler, M.;
2
(
(
(
(
6)
7)
8)
Stintzing, S.; Lenz, H. J. Clin Cancer Res 2014, 20, 3064.
Petroski, M. D. BMC Biochem 2008, 9 Suppl 1, S7.
Chen, D.; Frezza, M.; Schmitt, S.; Kanwar, J.; Dou, Q. P.
(
2
(
Curr. Cancer Drug Targets 2012, 11, 239.
9) Zavrski, I.; Jakob, C.; Kaiser, M.; Fleissner, C.; Heider, U.;
Sezer, O. Recent Results Cancer Res 2007, 176, 165.
10) Wang, Z.; Yang, J.; Kirk, C.; Fang, Y.; Alsina, M.; Badros,
(
(
(
A.; Papadopoulos, K.; Wong, A.; Woo, T.; Bomba, D.; Li, J.;
Infante, J. R. Drug Metab Dispos 2013, 41, 230.
(
1
(
(
(
Rademacher, J.; Heider, U.; Terpos, E.; Sezer, O. Expert Opin
Investig Drugs 2008, 17, 879.
(
(
E.; Mao, X.; Wang, X.; Zhang, Y.; Verbrugge, S. E.; Adomat, H.; Li,
Z. H.; Trudel, S.; Chen, C.; Religa, T. L.; Jamal, N.; Messner, H.;
Cloos, J.; Rose, D. R.; Navon, A.; Guns, E.; Batey, R. A.; Kay, L. E.;
Schimmer, A. D. J. Natl. Cancer Inst. 2010, 102, 1069.
11)
101.
12)
13)
14)
Beck, P.; Dubiella, C.; Groll, M. Biol Chem 2012, 393,
Kisselev, A. F. Chem Biol 2008, 15, 419.
Borissenko, L.; Groll, M. Chem. Rev. 2007, 107, 687.
Sterz, J.; von Metzler, I.; Hahne, J. C.; Lamottke, B.;
(
38)
de Wilt, L. H.; Jansen, G.; Assaraf, Y. G.; van Meerloo, J.;
Cloos, J.; Schimmer, A. D.; Chan, E. T.; Kirk, C. J.; Peters, G. J.;
Kruyt, F. A. Biochem. Pharmacol. 2012, 83, 207.
(
(
(
39)
40)
41)
Huber, E. M.; Heinemeyer, W.; Groll, M. Structure 2015.
Odom, A. L.; McDaniel, T. J. Acc Chem Res 2015.
Dissanayake, A. A.; Odom, A. L. Chem Commun (Camb)
15)
16)
Tobinai, K. Int J Clin Oncol 2007, 12, 318.
Mateos, M. V.; San Miguel, J. F. Best Pract Res Clin
2
(
012, 48, 440.
42) Swartz, D. L., 2nd; Staples, R. J.; Odom, A. L. Dalton
Trans 2011, 40, 7762.
Haematol 2007, 20, 701.
17) Kanagasabaphy, P.; Morgan, G. J.; Davies, F. E. Curr
Opin Investig Drugs 2007, 8, 447.
(
(
1
(
2
43)
1, 4720.
44)
001, 40, 1987.
45) Shi, Y.; Hall, C.; Ciszewski, J. T.; Cao, C.; Odom, A. L.
Chem Commun (Camb) 2003, 586.
Majumder, S.; Gipson, K. R.; Odom, A. L. Org Lett 2009,
(
7
(
1
(
8
18)
66.
19)
34, 253.
20)
53.
Barr, P.; Fisher, R.; Friedberg, J. Cancer Invest 2007, 25,
Vink, J.; Cloos, J.; Kaspers, G. J. Br J Haematol 2006,
Nau, K. C.; Lewis, W. D. Am Fam Physician 2008, 78,
Harris, S. A.; Ciszewski, J. T.; Odom, A. L. Inorg Chem
(

(
46)
Novak, A.; Blake, A. J.; Wilson, C.; Love, J. B. Chem
Commun (Camb) 2002, 2796.
(
3
(
47)
01, 3.
48)
New York, 1975.
49) Lee, S. W.; Kim, J. H.; Park, Y. B.; Lee, S. K. Ann.
Rheum. Dis. 2009, 68, 1761.
50) Nencioni, A.; Grunebach, F.; Patrone, F.; Ballestrero, A.;
Brossart, P. Leukemia 2007, 21, 30.
51) Tsukamoto, S.; Yokosawa, H. Expert Opin. Ther. Targets
009, 13, 605.
Gaczynska, M.; Osmulski, P. A. Methods Mol. Biol. 2005,
Segel, I. H. Enzyme Kinetics; John Wiley & Sons, Inc:
(
(
(
2
(
(
52)
53)
Napetschnig, J.; Wu, H. Annu Rev Biophys 2013, 42, 443.
Sors, A.; Jean-Louis, F.; Pellet, C.; Laroche, L.; Dubertret,
L.; Courtois, G.; Bachelez, H.; Michel, L. Blood 2006, 107, 2354.
54) Gokel, G. W.; Widera, R. P.; Weber, W. P. Organic
Syntheses 1976, 55, 232.
55) Li, A.-H.; Beard, D. J.; Coate, H.; Honda, A.; Kadalbajoo,
(
(
M.; Kleinberg, A.; Laufer, R.; Mulvihill, K. M.; Nigro, A.; Rastogi,
P.; Sherman, D.; Siu, K. W.; Steinig, A. G.; Wang, T.; D., W.; A.P.,
C.; Mulvihill, M. J. Synthesis 2010
1
678.